Compositions and methods to improve the homing and grafting of hematopoetic stem cells

Abstract

Methods comprising administering a composition including hematopoietic stem cells and one or more integrin activators or agonists, wherein the integrin activators or agonists improve homing, infiltrating, grafting, and retention of hematopoietic stem cells, especially, hematopoietic stem cells derived from umbilical cord blood, to reconstitute bone morrow tissue compromised by myeloablation therapy or by a hematologic cancer or a genetic disorder.

1 . A method comprising:
 administering a composition to a target tissue site of a patient, the composition comprising:
 a therapeutical amount of hematopoietic stem cells (HSCs), and 
 an effective amount of one or more integrin activating or agonist compounds, 
   wherein the effective amount is between about 1 fM and about 300 μM,   wherein the therapeutical amount of HSCs is about 10 6  cells, and   wherein the one or more integrin activating or agonist compounds enhance binding of surface integrins on the HSCs to their respective ligands and improve homing, infiltrating, grafting, and retention of HSCs to reconstitute bone morrow tissue compromised by myeloablation therapy or by a hematologic cancer or genetic disorder.   
     
     
         2 . The method of  claim 1 , further comprising:
 prior to administering step, treating a portion or all of the HSCs with one or more integrin activating or agonist compounds.   
     
     
         3 . The method of  claim 2 , further comprising:
 after the treating step, washing the treated HSCs to remove unbound integrin activating or agonist compounds.   
     
     
         4 . The method of  claim 1 , wherein, in the administering step, the cancer or genetic disorder is selected from the group consisting of acute myelogenous leukemia (AML), acute lymphoblastic leukemia (ALL), chronic myelogenous leukemia (CML), chronic lymphocytic leukemia (CLL), juvenile myelomonocytic leukemia, Hodgkin's lymphoma, non-Hodgkin's lymphoma, multiple myeloma, severe aplastic anemia, Fanconi's anemia, paroxysmal nocturnal hemoglobinuria (PNH), pure red cell aplasia, amegakaryocytosis/congenital thrombocytopenia, severe combined immunodeficiency syndrome (SCID), Wiskott-Aldrich syndrome, beta-thalassemia major, sickle cell disease, Hurler's syndrome, adrenoleukodystrophy, metachromatic leukodystrophy, myelodysplasia, refractory anemia, chronic myelomonocytic leukemia, agnogenic myeloid metaplasia, familial erythrophagocytic lymphohistiocytosis, solid tumors, chronic granulomatous disease, mucopolysaccharidoses, and Diamond Blackfan syndrome. 
     
     
         5 . The method of  claim 1 , wherein, in the administering step:
 the integrins are selected from the groups consisting of α4β1, α4β7, α5β1, αLβ2, αVβ3, and   mixtures or combinations thereof, and   the ligands are selected from the groups consisting of VCAM-1, fibronectin, MAdCAM-1, ICAM-1, ICAM-2, vitronectin, and mixtures or combinations thereof.   
     
     
         6 . The method of  claim 1 , wherein, in the administering step, the one or more integrin activating compounds comprise one or more compounds of the general Formula (I):
   R 1 -M 1 -N(R 2 )-M 2 -M 3 -M 4 -M 5 -M 6 -R 3   (I)
   
       wherein the compounds of Formula (I) is defined by:
 R 1  is selected from the group consisting of aryl and aralkyl, 
 R 2  is alkyl, aryl, or aralkyl, 
 M 1  is CH 2 , 
 M 2  is CO, 
 M 3  is O, S, or NR 6 , 
 R 6  when present is hydrogen or lower alkyl, 
 M 4  is absent or CH 2 , 
 M 5  is (CR 11 R 12 ),
 R 11  is hydrogen, 
 R 12  is selected from the group consisting of hydrogen, NR 21 CONR 22 R 23 , NR 21 COR 4 , NR 21 SO 2 R 24 , NR 21 COOR 24 , OCOR 24 , OR 24 , O(CH 2 CH 2 O) s R 24 , COOR 24 , alkyl, and hydroxyalkyl,
 s is an integer of 1 to 6, 
 
 R 21  and R 22  when present are independently selected from the group consisting of hydrogen or lower alkyl, 
 R 23  when present is selected from the group consisting of hydroxyalkyl, alkoxyalkyl, alkyl, aryl, aralkyl and alkoxycarbonylalkyl, 
 provided that when M 3  is NR 6  and M 4  is absent, then R 23  is not 1-(1,3-benzodioxol-5-yl)-3-ethoxy-3-oxopropyl, 
 R 24  when present is selected from the group consisting of alkyl, aryl, aralkyl, heterocyclyl, cycloalkyl, cycloalkylalkyl, and heterocyclylalkyl, and mixtures thereof, 
 
 M 6  is (CH 2 ) q , where
 q is an integer from 0 to 6, 
 
 R 3  is selected from the group consisting of hydrogen, CONR 13 R 14 , NR 15 COOR 16 , NR 15 COR 16 , NR 15 CONR 13 R 14 , NR 15 S02R 16 , OCOR 16 , COOR 16 , OR 16 , SR 16 , heterocyclyl, hydroxyl, hydroxyalkyl, guanadino, alkyl and aryl,
 R 13  and R 15  when present are independently hydrogen or lower alkyl, 
 R 14  and R 16  when present are independently selected from the group consisting of hydrogen, alkyl, aryl, aralkyl, cycloalkyl, heterocyclyl, cycloalkylalkyl, and heterocyclylalkyl, 
 
 R 1 , R 2 , R 3 , R 11 , R 14 , R 16 , R 23  and R 24  when present may independently be either unsubstituted or substituted with one or more substituents selected from the group consisting of alkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heterocyclylaryl, hydroxy, alkoxy, azido, haloalkoxy, hydroxyalkyl, aryloxy, hydroxyaryl, alkoxyaryl, halo, haloalkyl, haloaryl, amino, alkylamino, dialkylamino, arylamino, diarylamino, —NHCO(alkyl), —NHCO(aryl), —NHCO(aralkyl), —NHCO(haloalkyl), —NHSO 2 (alkyl), —NHSO 2 (aryl), —NHSO 2 (aralkyl), alkoxycarbonyl, alkoxycarbonylalkyl, —OCO(alkylamino), —OCO(dialkylamino), and mixtures thereof; 
 
       or
 pharmaceutically acceptable salts thereof, or mixtures thereof. 
 
     
     
         7 . The method of  claim 1 , wherein, in the administering step:
 R 1 M 1  and R 2  are independently selected from the group consisting of 2-thienylmethyl, 3-methoxybenzyl, 4-methoxybenzyl, 2-(2-thienyl)ethyl, pyridin-4-ylmethyl, 4-dimethylaminobenzyl, and pyridin-3-ylmethyl; and   when R 3  is CONR 13 R 14 , then R 13  and R 14  are independently selected from the group consisting of 2-thienylmethyl, 3-methoxybenzyl, 4-methoxybenzyl, 2-(2-thienyl)ethyl, pyridin-4-ylmethyl, 4-dimethylaminobenzyl, and pyridin-3-ylmethyl.   
     
     
         8 . The method composition of  claim 1 , wherein, in the administering step, the one or more integrin activating compounds comprise one or more compounds of the general Formula (I):
   R 1 -M 1 -N(R 2 )-M 2 -M 3 -M 4 -M 5 -M 6 -R 3   (I)
   
       wherein the compounds of Formula (I) is defined by:
 R 1  is aryl or aralkyl, 
 R 2  is alkyl or aralkyl, 
 M 1  is CH 2 , 
 M 2  is CO, 
 M 3  is absent or is O or CH 2 , 
 M 4  is absent or is CH 2 , 
 M 5  is absent or is O or (CR 11 R 12 ),
 R 11  is hydrogen, 
 R 12  is selected from the group consisting of hydrogen, NR 21 CONR 22 R 23 , NR 21 COR 24 , NR 21 SO 2 R 24  and NR 21 COOR 24 ,
 R 21  and R 22  each of which, when present is independently selected from the group of hydrogen and lower alkyl, 
 R 23  and R 24 , each of which, when present is independently selected from the group consisting of hydrogen, alkyl, aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl and aralkyl, 
 
 
 M 6  is selected from the group consisting of (CH 2 ) q , (CH 2 ) q —CH═CH—(CH 2 ) r , (CH 2 ) q -arylene-(CH 2 ) r , and (CH 2 CH 2 O) q , where
 q and r are independently integers from 0 to 6, 
 
 R 3  is CONR 13 R 14 ,
 R 13  and R 14 , each of which, when present is independently selected from the group consisting of hydrogen, alkyl, aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl and aralkyl, and 
 
 R 1 , R 2 , R 13 , R 14 , R 23  and R 24 , when present, independently either are unsubstituted or are substituted with one or more substituents selected from the group consisting of alkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heterocyclylaryl, hydroxy, alkoxy, azido, haloalkoxy, hydroxyalkyl, aryloxy, hydroxyaryl, alkoxyaryl, halo, haloalkyl, haloaryl, amino, alkylamino, dialkylamino, arylamino, diarylamino, —NHCO(alkyl), —NHCO(aryl), —NHCO(aralkyl), —NHCO(haloalkyl), —NHSO 2 (alkyl), —NHSO 2 (aryl), —NHSO 2 (aralkyl), alkoxycarbonyl, alkoxycarbonylalkyl, —OCO(alkylamino), —OCO(dialkylamino), and mixtures thereof; 
 
       or
 pharmaceutically acceptable salts thereof, or mixtures thereof. 
 
     
     
         9 . The method composition of  claim 1 , wherein, in the administering step:
 R 1 M 1  and R 2  are independently selected from the group consisting of 2-thienylmethyl, 3-methoxybenzyl, 4-methoxybenzyl, 2-(2-thienyl)ethyl, pyridin-4-ylmethyl, 4-dimethylaminobenzyl, and pyridin-3-ylmethyl; and   when R 3  is CONR 13 R 14 , then R 13  and R 14  are independently selected from the group consisting of 2-thienylmethyl, 3-methoxybenzyl, 4-methoxybenzyl, 2-(2-thienyl)ethyl, pyridin-4-ylmethyl, 4-dimethylaminobenzyl, and pyridin-3-ylmethyl.   
     
     
         10 . The method of  claim 1 , wherein, in the administering step, the one or more integrin activating compounds comprise one or more compounds of the general Formula (I):
   R 1 -M 1 -N(R 2 )-M 2 -M 3 -M 4 -M 5 -M 6 -R 3   (I)
   
       wherein the compounds of Formula (I) is defined by:
 R 1  is aryl or aralkyl, 
 R 2  is alkyl or aralkyl, 
 M 1  is CH 2 , 
 M 2  is SO 2  or CO, 
 M 3  is absent or is CH 2 , 
 M 4  is absent or is CH 2 , 
 M 5  is absent or is (CR 11 R 12 ),
 R 11 , when present, is hydrogen, 
 R 12 , when present, is selected from the group consisting of hydrogen, alkyl, NR 21 CONR 22 R 23 , NR 21 COR 24 , NR 21  SO2R 24  and NRC 21 COOR 24 ,
 R 21  and R 22 , each of which when present, is independently selected from the group of hydrogen, lower alkyl, and aralkyl, 
 R 23  and R 24 , each of which, when present is independently selected from the group consisting of hydrogen, alkyl, aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl and aralkyl, 
 
 
 M 6  is (CH 2 ) q , or NR 34 (CH 2 ) q ,
 q is an integer from 0 to 6, 
 R 34 , when present, is selected form the group consisting of alkyl, aralkyl, COR 35 , and SO 2 R 35 ,
 R 35  when present, is selected form the group consisting of alkyl, aryl, and aralkyl, and 
 
 
 R 3  is selected from the group consisting of CONR 13 R 14 , SO 2 NR 13 R 14 , NR 15 COOR 16 , NR 15 COR 16 , NR 15 CONR 13 R 14 , and NR 15 SO 2 R 16 ,
 R 13  and R 14 , each of which, when present, is independently selected from the group consisting of hydrogen, alkyl, aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl and aralkyl, 
 R 15  and R 16 , each of which when present, is independently selected from the group of hydrogen, lower alkyl, and aralkyl, 
 
 R 1 , R 2 , R 13 , R 14 , R 15 , R 16 , R 23 , R 24 , R 34  and R 35 , when present, either are unsubstituted or are substituted with one or more substituents selected from the group consisting of alkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heterocyclylaryl, hydroxy, alkoxy, azido, haloalkoxy, hydroxyalkyl, aryloxy, hydroxyaryl, alkoxyaryl, halo, haloalkyl, haloaryl, amino, alkylamino, dialkylamino, arylamino, diarylamino, —NHCO(alkyl), —NHCO(aryl), —NHCO(aralkyl), —NHCO(haloalkyl), —NHSO 2 (alkyl), —NHSO 2 (aryl), —NHSO 2 (aralkyl), alkoxycarbonyl, alkoxycarbonylalkyl, —OCO(alkylamino), and —OCO(dialkylamino), 
 with the proviso that when M 2  is CO, then M 6  is NR 34 (CH 2 ) q , q is not 0; 
 
       or
 pharmaceutically acceptable salts thereof, or mixtures thereof. 
 
     
     
         11 . The method of  claim 1 , wherein, in the administering step:
 R 1 M 1  and R 2  are independently selected from the group consisting of 2-thienylmethyl, 3-methoxybenzyl, 4-methoxybenzyl, 2-(2-thienyl)ethyl, pyridin-4-ylmethyl, 4-dimethylaminobenzyl, and pyridin-3-ylmethyl; and   when R 3  is CONR 13 R 14 , then R 13  and R 14  are independently selected from the group consisting of 2-thienylmethyl, 3-methoxybenzyl, 4-methoxybenzyl, 2-(2-thienyl)ethyl, pyridin-4-ylmethyl, 4-dimethylaminobenzyl, and pyridin-3-ylmethyl.   
     
     
         12 . The method of  claim 1 , wherein, in the administering step, the one or more integrin activating compounds comprise one or more compounds of the general Formula (I):
   R 1 -M 1 -N(R 2 )-M 2 -M 3 -M 4 -M 5 -M 6 -R 3   (I)
   
       wherein the compounds of Formula (I) is defined by:
 R 1  is alkyl, aryl or aralkyl, 
 R 2  is selected from the group consisting of aralkyl and alkyl,
 provided that when R 1  is alkyl, R 2  is aralkyl, 
 
 M 1  is CO or SO 2 ,
 provided that when M 1  is SO 2  and R 1  is phenyl, 4-methylphenyl or 2,4,6-trimethylphenyl, R 2  is not alkyl, 2-phenethyl, benzyl, or 2-methoxy-2-oxoethyl, and when M 1  is CO and R 1  is 2-furyl, 4-pyridyl, or 3,5-dinitrophenyl, R 2  is not alkyl, benzyl or 2-(1H-indol-2-yl)ethyl, 
 
 M 2  is absent or CH 2 , 
 M 3  and M 4  are absent, 
 M 5  is (CR 11 R 12 ),
 R 11  is hydrogen, 
 R 12  is selected from the group consisting of hydrogen, NR 2 CONR 22 R 23 , NR 2 COR 24 , NR 2 SO 2 R 24 , NR 21 COOR 24 , CONR 22 R 23 , COOR 24 , O(CH 2 CH 2 O), R 24 , hydroxyalkyl, and alkoxyalkyl,
 R 21 , and R 2 , when present, are independently selected from the group consisting of hydrogen and C 1 -C 6  alkyl, and 
 R 23  and R 24 , each of which, when present, is independently selected from the group consisting of hydrogen, alkyl, aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl and aralkyl, and
 s is an integer of 1 to 6, 
 
 
 
 M 6  is (CH 2 ) q ,
 q is an integer of 0 to 6, 
 
 R 3  is selected from the group consisting of NR 15 COOR 16 , NR 15 COR 16 , NR 15 CONR 13 R 14 , and 
 NR 15 SO 2 R 16 , and
 R 13  when present, is independently selected from the group consisting of hydrogen and C 1 -C 6  alkyl, and 
 R 14 , R 15 , and R 16  each of which, when present, are independently selected from the group consisting of hydrogen, alkyl, aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl and aralkyl, and 
 
 R 1 , R 2 , R 3 , R 12 , R 14 , R 15 , R 16 , R 23 , and R 24 , when present, independently either are unsubstituted or are substituted with one or more substituents selected from the group consisting of alkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heterocyclylaryl, hydroxy, alkoxy, azido, haloalkoxy, hydroxyalkyl, aryloxy, hydroxyaryl, alkoxyaryl, halo, haloalkyl, haloaryl, amino, alkylamino, dialkylamino, arylamino, diarylamino, —NHCO(alkyl), —NHCO(aryl), —NHCO(aralkyl), —NHCO(haloalkyl), —NHSO 2 (alkyl), —NHSO 2 (aryl), —NHSO 2 (aralkyl), alkoxycarbonyl, alkoxycarbonylalkyl, —OCO(alkylamino) and —OCO(dialkylamino), 
 
       or
 pharmaceutically acceptable salts thereof, or mixtures thereof. 
 
     
     
         13 . The method of  claim 1 , wherein, in the administering step:
 R 1 M 1  and R 2  are independently selected from the group consisting of 2-thienylmethyl, 3-methoxybenzyl, 4-methoxybenzyl, 2-(2-thienyl)ethyl, pyridin-4-ylmethyl, 4-dimethylaminobenzyl, and pyridin-3-ylmethyl; and   when R 3  is CONR 13 R 14 , then R 13  and R 14  are independently selected from the group consisting of 2-thienylmethyl, 3-methoxybenzyl, 4-methoxybenzyl, 2-(2-thienyl)ethyl, pyridin-4-ylmethyl, 4-dimethylaminobenzyl, and pyridin-3-ylmethyl.   
     
     
         14 . The method of  claim 1 , wherein the administering comprises parenteral injection. 
     
     
         15 . The method of  claim 14 , wherein the parenteral injection comprises intravenous injection, intra-arterial injection, or a combination thereof. 
     
     
         16 . The method of  claim 1 , wherein, in the administering step, the HSCs are derived from bone marrow, umbilical cord blood, peripheral blood, or mixture thereof. 
     
     
         17 . The method of  claim 1 , wherein, in the administering step, the HSCs are derived from bone marrow. 
     
     
         18 . The method of  claim 1 , wherein, in the administering step, the HSCs are derived from umbilical cord blood. 
     
     
         19 . The method of  claim 1 , wherein, in the administering step, the HSCs are derived from peripheral blood. 
     
     
         20 . The method of  claim 1 , wherein, in the administering step, the HSCs consist essentially of CD34 +  HSCs.